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利用超高孔 Ac-CGGASIKVAVS-OH 修饰的 PHEMA 支架促进人胎神经前体细胞的黏附和分化。

The use of superporous Ac-CGGASIKVAVS-OH-modified PHEMA scaffolds to promote cell adhesion and the differentiation of human fetal neural precursors.

机构信息

Institute of Experimental Medicine AS CR, vvi, Prague, Czech Republic.

出版信息

Biomaterials. 2010 Aug;31(23):5966-75. doi: 10.1016/j.biomaterials.2010.04.040. Epub 2010 May 18.

Abstract

Modifications of poly(2-hydroxyethyl methacrylate) (PHEMA) with laminin-derived Ac-CGGASIKVAVS-OH peptide sequences have been developed to construct scaffolds that promote cell adhesion and neural differentiation. Radical copolymerization of 2-hydroxyethyl methacrylate with 2-aminoethyl methacrylate (AEMA) and ethylene dimethacrylate in the presence of ammonium oxalate crystals resulted in the formation of superporous P(HEMA-AEMA) hydrogels. They were reacted with gamma-thiobutyrolactone to yield 2-(4-sulfanylbutanamido)ethyl methacrylate (P(HEMA-AEMA)-SH) unit. The Ac-CGGASIKVAVS-OH peptide was immobilized to the sulfhydryl groups of P(HEMA-AEMA)-SH by 2,2'-dithiodipyridine linking reagent via 2-[4-(2-pyridyldisulfanyl)butanamido]ethyl methacrylate (P(HEMA-AEMA)-TPy). The adhesion and morphology of rat mesenchymal stem cells were investigated on the Ac-CGGASIKVAVS-OH-modified P(HEMA-AEMA) as well as on PHEMA, P(HEMA-AEMA)-SH and P(HEMA-AEMA)-TPy hydrogels. Superporous Ac-CGGASIKVAVS-OH-modified PHEMA scaffolds significantly increased the number of attached cells and their growth area on the hydrogel surface in the absence and in the presence of serum in the culture medium. Additionally, the Ac-CGGASIKVAVS-OH peptide supported the attachment, proliferation, differentiation and process spreading of human fetal neural stem cells during the first two weeks of expansion and contributed to the formation of a high percentage of more mature neural cells after four weeks of expansion. The Ac-CGGASIKVAVS-OH modification of superporous P(HEMA-AEMA) hydrogels improves cell adhesive properties and promotes neural stem cell differentiation.

摘要

聚(2-羟乙基甲基丙烯酸酯)(PHEMA)与层粘连蛋白衍生的 Ac-CGGASIKVAVS-OH 肽序列的修饰已被开发用于构建促进细胞黏附和神经分化的支架。在草酸铵晶体存在下,将 2-羟乙基甲基丙烯酸酯与 2-氨基乙基甲基丙烯酸酯(AEMA)和二乙烯基甲醚自由基共聚,形成超多孔 P(HEMA-AEMA)水凝胶。它们与 γ-巯基丁内酯反应生成 2-(4-磺基丁酰胺基)乙基甲基丙烯酸酯(P(HEMA-AEMA)-SH)单元。Ac-CGGASIKVAVS-OH 肽通过 2,2'-二硫代二吡啶连接试剂固定在 P(HEMA-AEMA)-SH 的巯基上,生成 2-[4-(2-吡啶基二硫基)丁酰胺基]乙基甲基丙烯酸酯(P(HEMA-AEMA)-TPy)。在 Ac-CGGASIKVAVS-OH 修饰的 P(HEMA-AEMA)以及 PHEMA、P(HEMA-AEMA)-SH 和 P(HEMA-AEMA)-TPy 水凝胶上研究了大鼠间充质干细胞的黏附和形态。超多孔 Ac-CGGASIKVAVS-OH 修饰的 PHEMA 支架在培养基中存在和不存在血清的情况下,显著增加了附着细胞的数量及其在水凝胶表面上的生长面积。此外,Ac-CGGASIKVAVS-OH 肽支持人胎神经干细胞在扩增的头两周的附着、增殖、分化和突起伸展,并有助于在扩增四周后形成更高比例的更成熟的神经细胞。超多孔 P(HEMA-AEMA)水凝胶的 Ac-CGGASIKVAVS-OH 修饰改善了细胞黏附性能,并促进了神经干细胞的分化。

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